Thalassemic Subjects Research Articles

Clinical characteristics, laboratory features and genetic profile of hemoglobin E (HBB:c.79 G > A)/β (nucleotide -28 A > G) (HBB:c.-78 A > G) -thalassemia subjects identified from community- and hospital-recruited cohorts

Despite several existing laboratory-based studies of hemoglobin (Hb) E (HBB:c.79 G > A)/ β (nucleotide (NT) -28 A > G) (HBB:c.-78 A > G) -thalassemia, no reports have ever provided clinical severity information as well as dependency of blood transfusion. Previously, a comparative study of community- and hospital-recruited Hb E/β-thalassemia subjects was conducted in the lower northern Thailand between June 2020 and December 2021. A mobile medical team visited each community hospital on-site, collecting clinical severity parameters, and conducting Hb and DNA analyses. The control included Hb E/β-thalassemia patients undergoing transfusions. Subgroup study of adult Hb E/β (NT -28 A > G) -thalassemia subjects was subsequently conducted. Additional pediatric individuals were recruited from prenatal diagnosis databases. Twenty adult and nine pediatric subjects were enrolled; all were classified as having mild disease severity. Twenty-two individuals (75.9 %) were asymptomatic. Six adults (20.7 %) required blood transfusion. The mean Hb level of subjects without transfusion (23 [79.3 %]) was 10.77 ± 1.10 g/dL. Hb analysis revealed a distinct EFA pattern with low Hb F fraction. The positive impact of genetic modifiers could not be statistically demonstrated except rs7482144-XmnI. These findings could provide essential information for parents carrying fetuses with Hb E/β (NT -28 A > G) -thalassemia.

Testicular Volumes Measurement Comparison by Ultrasound and Orchidometer and Its Relationship with Nutritional Status in Transfusion-dependent Thalassemia

BACKGROUND: One type of thalassemia known as transfusion-dependent thalassemia (TDT) requires a lifelong blood transfusion. Iron overload from prolonged blood transfusions causes nutritional disorders and several organs impairment, including the testicles. In contrast to an orchidometer, ultrasound shows more details of the testicles. This study aimed to compare testicular volumes based on ultrasound and orchidometer measurements and correlate testicular volume with nutritional status in TDT patients.METHODS: A cross-sectional study including 27 male thalassemia subjects who underwent routine check-ups was conducted. Measurements of testicular volume using an orchidometer and ultrasound, and body mass index (BMI) was performed. Ultrasound measurement of testicular volume was calculated using the Lambert Formula, while the orchidometer was performed by matching the bead next to the testicle.RESULTS: Most subjects were underweight (63%) or normal weight (37%). However, no significant correlation between BMI and testicular volume was found. As measured using an orchidometer, the mean volume of the right and left testes were 9.22±4.92 mL and 8.67±4.45 mL, respectively. Ultrasound measurement showed the mean values of the right and left testicles were 7.68±3.96 mL and 7.09±4.03 mL, respectively. Testicular measurements obtained using ultrasound showed a significant difference (p<0.05) when compared with the orchidometer. The testicle measured by ultrasound was smaller, which was more accurate. However, the comparison between the right and the left testicular volumes showed no significance.CONCLUSION: Thalassemia patients are mainly underweight, and their nutritional status have no relationship with testicular volume. Ultrasound, as compared to an orchidometer, is better for measuring testicular volume.KEYWORDS: thalassemia, testicle, orchidometer, ultrasound, body mass index

Early Effects of a Repurposed Therapeutic PB-04 in Beta Thalassemia Intermedia and in Transgenic Sickle Mice

Background: Biochemical, epidemiologic, genetic, and clinical data over 7 decades have shown that renewed or persistent expression of fetal globin reduces clinical severity in beta-thalassemias (BTI) and hemoglobinopathies (sickle cell disease, SCD). Hydroxyurea (HU) and epigenetic therapies which act through HDAC and DNMT-1 inhibition have demonstrated beneficial clinical activity; butyrate, a Class I HDAC inhibitor which suppresses BCL11A, increased HbF by 3-fold and total hemoglobin (Hb) by 1 gm/dl in SCD and eliminated transfusions in beta thalassemia patients, and decitabine strongly induces HbF, increases total Hb, and reduces biomarkers of sickling. Active erythropoiesis, basal EPO levels, available iron, and select mutations or modifiers are associated with responses. Additional small molecule therapies, especially those with additive activity with HU, are considered desirable for many patients world-wide. Methods: Seeking oral noncytotoxic therapeutic candidates with low development risks, we utilized a high-throughput screen with a gamma globin promoter-reporter and cytotoxicity readout for unrecognized fetal globin-inducing activity in libraries of all FDA- and EMA-approved or clinical stage therapeutics (for any medical indication). PB-04, a candidate with a benign long-term safety profile (used to inhibit L-dopa metabolism in Parkinson's disease), does not inhibit proliferation of erythroid progenitors and suppresses or displaces 4 repressors of the fetal globin gene promoter, BCL11A, HDAC3, KLF1, LSD-1, indicating targeted molecular activity. PB-04 has additive effects with HU in inducing F-cells in sickle cell erythroid progenitors. PB-04 was evaluated in 3 predictive animal models and in 3 escalating dose cohorts in adult subjects with BTI. Results: In 3 animal models, PB-04 increased fetal globin mRNA by 12-to 20-fold and F-cells by 20% in anemic baboons, F-cells and mean Hb F/cell by 2 to 7-fold in beta-YAC mice, and fetal globin mRNA by 1.8-fold and % F-cells by 2.4-fold without toxicity in Townes SCD mice, similar to the changes induced by HU (both given 5 days/wk for 4 wks). In six beta thalassemia (BTI) subjects who completed ten 12-week courses at 1, 3, or 5 mg/kg doses given once/day, 3 times/wk, increases from baseline in F-reticulocytes (mean 5.2-fold, range 1.5-15.7), F-cells, (mean 4.4-fold, range 2.3 to 7.3-fold), and HbF/cell (MFI), (mean 6.6-fold, range 3 to 11.3-fold), were observed (p<0.03 for all parameters). Increases were observed even in subjects with low baseline HbF levels. HbF increases by HPLC were 2.7 to 14%. Treatment emergent adverse effects (TEAEs) have been Gr 1-2 and largely transient in these subjects receiving 12 weeks' therapy. Other dosing regimens are being investigated. Conclusions: Initial studies in beta globin disorders indicate PB-04 induces fetal globin protein expression assayed by F-reticulocytes, F-cells, and mean HbF/cell in BTI patients and in transgenic SCD mice without toxicity. These findings indicate PB-04, which has complementary activity to all the approved SCD therapeutics, merits clinical evaluation in patients with sickle cell disease.

Symphysis morphology and mandibular alveolar bone thickness in patients with β-thalassemia major and different growth patterns

ABSTRACTObjective: The present study aimed to assess the morphology of symphysis and alveolar bone thickness (ABT) surrounding mandibular incisors in thalassemic patients, as compared to unaffected individuals. Methods: This case-control study was conducted on lateral cephalograms of 60 thalassemic and 60 unaffected patients with Class II malocclusion seeking orthodontic treatment at Dental School, Shiraz University of Medical Sciences. The sample was divided into three subgroups including hyperdivergent, normodivergent, and hypodivergent, according to the Jarabak index. Symphysis dimensions and alveolar bone thickness surrounding mandibular incisors were measured using AutoCad software. Finally, the correlation between alveolar bone thickness and symphysis morphology was assessed. Results: In general, chin dimensions and bone thickness at different levels of mandibular incisor roots (cervical, middle, apical) were smaller in thalassemic adolescents than controls. Concerning the total sample as well as the normodivergent subgroup, significantly lower values were observed in thalassemic patients for symphysis width, total ABT at the cervical, and lingual ABT at the apical root area compared to controls (p < 0.05). The hypodivergent growth pattern was not associated with any statistical differences between the groups (p> 0.05). In both thalassemic and control subjects, symphysis width showed a weak to moderate positive correlation with ABT of lower incisors (p< 0.05), whereas symphysis height showed a moderate positive correlation with cervical ABT in only ß‐thalassemia patients (p< 0.05). Conclusions: Compared to controls, ß-thalassemia patients showed thinner alveolar bone at different levels of lower incisor roots and smaller symphysis dimensions. There were significant correlations between symphysis dimensions and alveolar bone thickness of mandibular incisors in the sample.

Uric Acid Abnormalities and Its Correlation with Splenectomy in Adult Transfusion Dependent Thalassemia Subjects (TDT)

Background: In thalassemia patients erythrocyte turnover rate increases due to chronic hemolysis and ineffective erythropoiesis leading to increase in uric acid production. Hyperuricosuria is one of the marker of proximal tubular dysfunction. Splenectomy can increase the risk of hyperuricemia by increasing erythrocyte turnover rate in Transfusion Dependent Thalassemia (TDT). Deferasirox enhances uric acid excretion from renal tubules leading to low serum uric acid levels. Methods: An institution based cross-sectional study was conducted from November 2018-March 2020 which included 60 adult TDT patients (≥18 years) attending Adult Thalassemia Day Care Centre at tertiary care hospital. Serum uric acid (mg/dl), 24 hour urinary uric acid (mg/dl), serum ferritin (ng/ml) were done. Results: Hyperuricemia was present in 20%, hypouricemia in 3.33%. Hyperuricosuria was present in 80%. Mean SUA levels were higher in males than females which was statistically significant (5.77±1.66 vs 4.64±1.44, p=0.007). There was statistically significant positive correlation between serum uric acid and serum creatinine(p = 0.0036) and age (p=0.0129). Hyperuricemia was more in subjects with intact spleen but was not statistically significant(p=0.104). Hyperuricemia had negative association with deferasirox therapy [1 (3.03%) vs 11 (40.74%), p=0.0004] compared with normouricemia subjects. Conclusion: Uric acid abnormalities were seen in 23.3% of patients. Hyperuricemia was present in one fourth of the patients and significant risk factors were increasing age, male gender, rising serum creatinine. Deferasirox therapy was negatively associated with hyperuricemia. Tubular dysfunction is relatively common in TDT as more than three fourth patients had hyperuricosuria. Regular monitoring of serum uric acid and urinary uric acid is recommended.

Estimation of serum ferritin and TSH levels in thalassemia patients undergoing iron chelation therapy

Objectives &amp; introduction: Thalassemia, heterogenous group of disorders of haemoglobin,characterised by reduced or absent production of one or more of globin chains.Regular redcell transfusion with chelation therapy for iron overload are cornerstones of therapy for ßthalassemia major. Serum ferritin assay is widely available, relatively inexpensive methodfor assessing body iron burden and monitoring response to chelation process which in turnalso improves TSH levels in thalassemic subjects .The objective of this study was to assessprechelation and postchelation levels of serum ferritin and TSH and correlating post chelationlevels of serum ferritin and TSH in thalassemic patients &gt;6yrs undergoing chelation therapy.&#x0D; Materials &amp; methods: Serum TSH measured by Enzyme linked fluorescent assay and serumferritin measured by enzyme linked immunosorbent assay.&#x0D; Results: Amongst 500 participants,47% were males &amp; 53% females. Mean age was 9.04 yrs ;prechelation ferritin and TSH levelswere 2995.78ng/ml with SD of 802.53 and 5.07 μU/ml with SD of 2.52. The postchelationferritin and TSH levels were 2168.80 ng/ml with SD of 1335.89 and 4.51μU/ml with SD of4.76. Paired t test with respect to pre and postchelation ferritin and TSH levels showed 2 tailed pas 0.000 and t&gt;3, both of which considered significant. While correlating post chelation ferritinwith TSH levels; they showed a linear correlation ( Pearson coefficient of .836).&#x0D; Conclusion:Serum ferritin and TSH estimation in prechelation and postchelation periods give an estimateof iron overload with effect of chelation on it. Both levels decrease post chelation presenting alinear correlation between the two.&#x0D; Bangladesh Journal of Medical Science Vol.20(1) 2021 p.130-135

COGNITIVE IMPAIRMENT DETECTION IN ADULT THALASSEMIA PATIENT USING MOCA-INA

Background: Cognitive impairment in thalassemia patients are prevalent, therefore early detection of cognitive impairment in adult thalassemia patients is crucial for prevention. Montreal Cognitive Assessment (MoCA) is a public domain cognition screening tools that covers all cognitive domains in detecting mild cognitive impairments. Objective: To compare cognitive function between adult thalassemia patients and healty control by using Indonesia version of MoCA test (MoCA-Ina) Methods: This prospective observational analytic with case control study, compared the total scores and scores of each domain of cognition between adult thalassemia patients and healthy subjects at the Medical Hematology Oncology Clinic of Dr. Hasan Sadikin General Hospital, Bandung, Indonesia using MoCA-Ina from August to October 2018. Results: A total of 32 thalassemia subjects and 50 healthy subjects were conducted. A total of 16(50%) subjects in the case group had a value of MoCA-Ina &lt;26, while only 1(2%) healthy control had a value of MoCA-Ina &lt;26. The median total MoCA-Ina score in case and control groups were 25.5 and 27.50 (p &lt;0.001). The median score of memory domains, executive functions and visuospatial of the case and control groups were 3 versus 4 (p &lt;0.001), 3 versus 3.5 (p &lt;0.001) and 3.53 ± 0.671 versus 3.88 ± 0.385 (p &lt;0.003), respectively. Conclusion: Adults thalassemia patients have lower score in total MoCA-Ina, domains of memory, executive function and visuospatial score compared to healthy control.

High Prevalence of Antiphospholipid Antibodies in Children with Non-Transfusion Dependent Thalassemia and Possible Correlations with Microparticles.

Antiphospholipid antibodies (APAs) are developed antibodies to the phospholipid surfaces or to the proteins binding to phospholipids. Increased PS-expressed red blood cells (RBC) may be a risk factor in APAs development. There were only a few studies reported on APAs in thalassemia, and their associations to PS-expressing microparticles (MPs), remain unknown. The presence of antiphospholipid antibodies (APAs) in pediatric thalassemia patients and their associations with phosphatidylserine (PS)- expressing microparticles (MPs), a previously unexplored relationship, were the aims of this report. We identified that positive APA rates were highest in non-transfusion-dependent thalassemia (NTDT) subjects, followed by transfusion-dependent thalassemia (TDT) subjects and controls (29.5%, 18.2% and 17.9%, respectively). Subgroup analysis demonstrated that positive anti- ?2 glycoprotein I (a?2-GPI)- IgG rates in NTDT subjects were significantly greater than in controls and that the percentage of MPs was also significantly elevated in thalassemia subjects. More specifically, there were significant correlations found between a?2GPI-IgG and anticardiolipin (aCL)-IgM APAs, and MPs.

Assessment of mandibular characteristics in patients affected with β-thalassaemia major: A retrospective case-control study

Thalassemia is the most common hereditary blood disorder across the world. This study aimed to identify some mandibular features of thalassaemic patients and compare them with unaffected counterparts. This retrospective case-control study was carried out on lateral cephalograms of 60 subjects (26 males, 34 females) with class II malocclusion and age range of 11 to 15 years. The control group consisted of 60 non-thalassaemic subjects with class II malocclusion and similar chronological age, gender and vertical facial dimension. Based on the Jarabak index, the case and control subjects were classified into hyperdivergent, normodivergent and hypodivergent growth patterns. Four linear (ramus height, ramus width, mandibular depth, and antegonial notch depth) and 3 angular (symphyseal angle, gonial angle, and mandibular arc angle) cephalometric parameters were measured to represent mandibular morphology. The data were analysed using Chi-square test and Student's t-test. No significant difference was found in linear measurements between thalassaemic patients and controls. The symphysis angle was significantly greater and the mandibular arc angle was significantly smaller in the total thalassaemic sample than the control individuals (P<0.001 and P=0.004, respectively). The difference in symphysis angle was significant in both hyperdivergent and normodivergent subjects (P=0.004 and P=0.002, respectively), whereas the difference in mandibular arc angle was only significant in the normodivergent subgroup P=0.001). The smaller mandibular arc angle in the thalassaemic sample suggests a more superior than posterior growth direction of condyles compared with healthy individuals. The difference in symphyseal angle represents inherent differences in chin morphology between thalassaemic subjects and controls.

An Innovative Multilevel Test for Hemoglobinopathies: TGA/Chemometrics Simultaneously Identifies and Classifies Sickle Cell Disease From Thalassemia.

Introduction: Hemoglobinopathies are the most common genetic disorder wordlwide and because of migrations are become an emerging global health problem. Screening programmes for Sickle cell disease and Thalassemia have been implemented in some countries, but are not a common practice, due to a lack in the accuracy of the methods and to the costs of the analyses.Objectives: The objective of this study was the application of the thermogravimetry coupled to chemometrics as new screening method to perform an early diagnosis of thalassemia and sickle cell disease.Methods: Whole blood samples (30 μL) from sickle cell anemia and thalassemia patients were analyzed using the thermobalance TG7 and the resulting curves were compared with those of healthy individuals. A chemometric approach based on Principal Components Analysis (PCA) was exploited to enhance correlation between thermogravimetric profiles and a model of prediction by Partial Least Square Discriminant Analysis (PLS-DA) was developed and validated.Results: The characteristic profile of the blood sample thermal decomposition and the first derivative of the TG curve showed that patients were clearly distinguished from healthy individuals as a result of different amounts of water and corpuscular fraction of blood. The chemometric approach based on PCA allowed a quick identification of differences between healthy subjects and patients and also between thalassemic and sickle cell anemia subjects. Chemometric tools (PLS-DA) were used to validate a model of prediction to process the thermogravimetric curves and to obtain in 1 h an accurate diagnosis. The TGA/Chemometric test permitted to perform first level test for hemoglobinopathies with the same accuracy of confirmatory analyses obtained by the molecular investigation.Conclusions: A screening test based on the coupling of thermogravimetry and chemometrics was optimized for the differential diagnosis of hemoglobinopathies. The novel test is able to simultaneously perform a simple and fast diagnosis of sickle cell anemia or thalassemia, in a single analysis of few microliters of non-pretreated whole blood at low cost, and with high accuracy. Moreover this method results particularly suitable in pediatric patients as it requires small sample volumes and is able to characterize also transfused patients.

Red cell alloantibodies in beta-thalassaemia major patients' blood referring to the regional blood transfusion center of Tehran, Iran.

Introduction: Thalassemia is associated with a genetic decline in the rate of synthesis of one or more types of natural hemoglobin polypeptide chains. One of the major complications in thalassemia patients is alloimmunization, which is antibody production by the patient against transfused red blood cells (RBCs). These RBCs are unknown by the recipient and the formed antibodies against them are called alloantibodies. This study aimed to evaluate the frequency of alloantibodies against RBCs in beta-thalassemia patients referred to Tehran Regional Blood Transfusion Center. Methods: In this study, antibody screening tests (Dia-cell I, II, and III) were performed on 184 thalassemia patients. An identification test by the Dia panel consisting of 11 different O RBCs groups to examine sera with Dia cells (I, II, or III) was performed. Results: In our study, males and females patients comprised 66 (35.87%) and 118 (64.13%), respectively, of whom 116 (63%) had alloimmunization. In addition, 68 thalassemia subjects (37%) lacked alloantibodies. Among 184 patients with beta-thalassemia major, anti-K (Kell system), anti-D, and anti-E (Rhesus system) had the most abundant alloantibody variants with an incidence of 24 (13%), 11 (5.98%), and 10 (5.4%), respectively. Conclusion: Before RBC transfusion, regular RBC antigen phenotypes, as well as problem-solving of alloantibody production by receiving compatible blood for Kell and RH subgroups, are suggested for all cases of transfusion-derived thalassemia.

High Tissue Factor Microparticle Level in Major Thalassemic Patients with Pulmonary Hypertension

Objective: To analyze the correlation between tissue factor microparticles (TF-MP) levels and pulmonary hypertension (PH) in adult thalassemic patients. Methods: This study was conducted from September to October 2018, using secondary and primary data. The secondary data consisted of the PH parameter, which was retrieved from a 2017 previous study entitled ‘Clinical Characteristic and Complication due to Iron Overload in Thalassaemic Patients‘in 2017 while the primary data were the TF-MP, which were obtained from the analysis of frozen serum of the same population using ELISA method. The mean pulmonary arterial pressure (mPAP) values were obtained from echocardiography results and PH was defined as mPAP &gt;25 mmHg. Results: Seven (16.7%) major thalassemic patients experienced PH. The median values of TF-MP levels were higher among major thalassemic patients with PH when compared to the non-PH patients (1569 vs 11.5 pg/dL; p=0.023). No significant difference was observed in the median TF-MP levels between subjects with splenectomy and subjects without splenectomy (11.6 vs 12.3 pg/dL; p=0.44). There was also no difference in mPAP values between subjects with splenectomy and subjects without splenectomy (18.0 vs 17.0 mmHg; p=0.663). When the median TF-MP levels among major thalassemic patients were analyzed in terms of correlation with transfusion level, no statistically significant difference was seen between subjects who received sufficient transfusions (≥180 mL/kgbb/year) and those who received insufficient transfusions (&lt;180 mL/kgbb/year) (r= 0.138; p=0.390). Conclusions: There is a positive correlation between the TF-MP levels and PH in adult major thalassemic subjects.

Safety Study for Beta Thalassemia Subjects on PTG-300

Safety Study for Beta Thalassemia Subjects on PTG-300

Immunostick Test for Detecting \u03b6-Globin Chains and Screening of the Southeast Asian \u03b1-Thalassemia 1 Deletion

BackgroundCouples who carry α-thalassemia-1 deletion are at 25% risk of having a fetus with hemoglobin Bart’s hydrops fetalis. Southeast Asian deletion (−-(SEA)) is the most common type of α-thalassemia 1 among Southeast Asian populations. Thus, identification of the (−-(SEA)) α-thalassemia 1 carrier is necessary for controlling severe α-thalassemia in Southeast Asian countries.ResultsUsing our generated anti ζ-globin chain monoclonal antibodies (mAbs) clones PL2 and PL3, a simple immunostick test for detecting ζ-globin chain presence in whole blood lysates was developed. The procedure of the developed immunostick test was as follows. The immunostick paddles were coated with 50 μg/mL of mAb PL2 as capture mAb, or other control antibodies. The coated immunostick was dipped into cocktail containing tested hemolysate at dilution of 1:500, 0.25 μg/mL biotin-labeled mAb PL3 and horseradish peroxidase-conjugated streptavidin at dilution of 1:1000. The immunostick was then dipped in precipitating substrate and the presence of ζ-globin chain in the tested sample was observed by the naked eye. Upon validation of the developed immunostick test with various types of thalassemia and normal subjects, 100% sensitivity and 82% specificity for detection of the (−-(SEA)) α-thalassemia-1 carriers were achieved. The mAb pre-coated immunostick can be stored at room temperature for at least 20 weeks.ConclusionIn this study, a novel simple immunostick test for the screening of (−-(SEA)) α-thalassemia 1 carriers was presented. The developed immunostick test, within a single test, contains both positive and negative internal procedural controls.

Cellular adaptation mediated through Nrf2-induced glutamate cysteine ligase up-regulation against oxidative stress caused by iron overload in β-thalassemia/HbE patients

Oxidative stress caused as a result of iron overload is implicated in clinical manifestation of beta-thalassemia/haemoglobin E (β-Thal/HbE). In this study, we investigated the cellular adaptation against oxidative stress in β-Thal/HbE patients. Twenty-four paediatric β-Thal/HbE patients and 22 healthy controls were recruited in the study. Blood samples from patients exhibited iron overload, elevation of lipid peroxidation, and marked diminution in the reduced glutathione (GSH) level. However, expression of glutamate-cysteine ligase catalytic (GCLC) subunit, a key enzyme in GSH biosynthesis, was up-regulated when compared with that in controls. GCLC protein levels were correlated with serum iron. There was an enhanced binding activity of the oligonucleotide probe for Nrf2-driven antioxidant response element (ARE) to nuclear protein from blood mononuclear cells of thalassemia subjects. In conclusion, β-Thal/HbE patients exhibit elevated plasma levels of GCLC expression and Nrf2-ARE binding activity, which may account for their adaptive survival response to oxidative stress.

PB2392 GLOBAL HEMOSTATIC CHANGES AND CIRCULATING MICROPARTICLES IN CHILDREN AND ADOLESCENTS WITH HEMOGLOBIN H AND BETA‐THALASSEMIA/HEMOGLOBIN E DISEASES

Background:Thalassemia is an inherited hemolytic anemia known to be associated with hypercoagulable state. Several factors including the abnormality of thalassemic red blood cell (RBC) membrane, increased platelet count, decreased anticoagulant proteins and increased circulating microparticles (MPs) contribute to the hypercoagulability in thalassemia. Global hemostatic changes as measured by rotatory thromboelastometry (ROTEM®) have been demonstrated in patients with beta‐thalassemia/hemoglobin E disease (beta‐thal/Hb E). However, there is little information in patients with hemoglobin H disease (Hb H) which is a less severe but highly prevalent non‐transfusion dependent thalassemia.Aims:To compare the global hemostatic changes and the levels of circulating MPs among children and adolescents with Hb H, beta‐thal/Hb E and healthy controls.Methods:A cross‐sectional study was conducted in 38 subjects with Hb H, 22 subjects with beta‐thal/Hb E and 12 healthy age‐matched controls. Complete blood count, ROTEM® and quantitation of circulating MPs [annexin V + MPs, red blood cell (RBC) derived MPs (RMPs) and platelet derived MPs (PMPs)] as analyzed by flow cytometry were performed in all subjects.Results:The age (range 5‐21 years) and percentage of male gender (range 50‐58%) among the Hb H, beta‐thal/Hb E and control groups were not statistically different (P = 0.383 and P = 0.798, respectively). The Hb levels were higher and the numbers of nucleated RBC (nRBC) were lower in the Hb H group than in beta‐thal/Hb E group. Overall, the ROTEM® parameters were not statistically different among the thalassemia and control groups. However, within the thalassemia subjects group, the maximum clot firmness (MCF) in splenectomized patients was significantly higher than non‐splenectomized subjects (P &lt; 0.001). The levels of annexin V + MPs in both Hb H and beta‐thal/Hb E groups were significantly higher than the control group (P = 0.001). RMPs in beta‐thal/Hb E group were higher than the controls (P = 0.013) while there was no difference between the Hb H and control groups (P = 0.097). The level of circulating MPs was slightly higher in splenectomized subjects, but the differences were not statistically significant. Only the platelet count and nRBC, but not the circulating MPs, were significantly correlated with MCF in both INTEM (r = 0.620, P &lt; 0.001 and r = 0.322, P = 0.006 respectively) and EXTEM (r = 0.567, P &lt; 0.001 and r = 0.326, P = 0.005, respectively).Summary/Conclusion:The global hemostasis parameters as measured by ROTEM® were not different among the Hb H, beta‐thal/Hb E and control groups in this study. Hypercoagulable state was more commonly found in thalassemia subjects who were splenectomized, regardless of genotypes. The hypercoagulability was associated with higher nRBC and platelet count in the splenectomized group. Patients with Hb H had significantly higher circulating MPs levels than healthy controls although with the lesser extent than those seen in beta‐thal/Hb E.

New methods for thalassemia screening: TGA/Chemometrics test is not influenced by the aging of blood samples

The coupling of Thermogravimetry (TG) in conjuction with chemometrics was investigated for the first time to evaluate the capabilities of this novel test to provide the screening of thalassemia in blood samples stored at 4 °C until 15 days. Healthy donors were considered as reference subjects and a typical thermal behaviour as a function of aging was estimated and compared to thermal behaviour of thalassemia subjects. Diagnosis of thalassemia was made at the Day-Hospital Thalassemia of S. Eugenio Hospital through a comprehensive assessment of clinical presentation and hematological and molecular analysis. Despite blood changes with aging, the application of the TGA/Chemometrics test reveal that healthy and thalassemic population may be significantly differentiated after 15 days from blood collection with a 100% of correct classification rate. This new method applied to aged samples was able to discriminate thalassemia in transfused patients that is generally not possible by the common first level protocol used for thalassemia screening, and in δβ-thalassemias, and β-thalassemia combined with Hb Lepore, usually requiring the molecular analysis for diagnosis. This study, for the first time, describes a screening method for thalassemia able to detect thalassemia on whole blood samples stored for 15 days.In conclusion TGA/Chemometrics screening test for thalassemia is not influenced by the aging of blood samples and this approach could open the way to prevention programs for thalassemia also in the developing countries where expertise and facilities for their control are extremely limited or in situation involving delayed analyses.

Divergent Pro-Inflammatory Cytokine Response Induced by GB Virus C/ Hepatitis G virus (GBV-C/HGV) and Torque Teno Virus (TTV) Co-Infections in Hepatitis C Virus (HCV) Infected Thalassemia Patients

Despite common incidence of GB virus C/Hepatitis G virus (GBV-C/HGV) and Torque Teno Virus (TTV) co-infections in subjects with Hepatitis C virus (HCV) infection, pathogenic role of these co-infections in HCV infected subjects has not been clear since studies have mostly been based on liver enzyme profile yielding variable results. Pro-inflammatory cytokines that participate in host immune response against viruses are generated as a consequence of triggering of related genes by Nuclear Factor Kappa B (NF-kB) that is a member of transcription family. Study of three pro-inflammatory cytokines e.g. interleukin 1 beta (IL-1&amp;beta;), Interleukin-6 (IL-6), interleukin-8 (IL-8) in a group of multi-transfused thalassemic subjects in relation to positivity for HCV, HGV and TTV infections showed elevated levels of these cytokines in serum as well as in supernatant of peripheral blood mononuclear cell (PBMCs) cultures in HCV-GBV-C/HGV co-infected subgroup compared to HCV mono-infected subgroup (p &amp;lt;0.05 for comparison of all three cytokines) while HCV-TTV co-infected subgroup showed lowering of these levels compared to HCV mono-infected subgroup (p &amp;lt;0.05 for comparison of all three cytokines). Levels of p65 component of NF-kB i.e.NF-kB p65 in nuclear extracts of lipopolysaccharide stimulated PBMCs correlated positively with the levels of pro-inflammatory cytokines in serum as well as that in supernatants of PBMC cultures in both HCV- GBV-C/HGV co-infected and HCV-TTV co-infected subgroups (p values ranging from &amp;lt;0.001 to 0.004). Levels of NF-kB p65 in nuclear extracts and levels of pro-inflammatory cytokines in serum as well as in supernatants of PBMC culture did not show any alteration compared to thalassemic subjects with HGV or TTV infections alone and healthy non-transfused subjects. Based on the inhibitory role of TTV on activation of NF-kB in TTV-HCV co-infected cases observed in the present study and the reported contribution of NF-kB towards development of hepatocellular carcinoma (HCC) due to establishment of chronicity, it may be worth evaluating if TTV or any component of TTV can be utilized as therapeutic vaccine against development of HCC in HCV infected subjects.

Tooth Crown Size of the Permanent Dentition in Subjects with Thalassemia Major

Thalassemia refers to a group of hereditary anemias resulting from defects in synthesis of either alpha or the beta polypeptide chains of hemoglobin. The homozygous form of beta thalassemia (thalassemia major) exhibits the most severe clinical symptoms. Odontometric analysis of subjects with thalassemia are lacking, despite the wide distribution of the disease and its orofacial characteristics. The aim of this study was to provide detailed descriptions of tooth crown size in the permanent dentition of subjects with thalassemia major and to compare the findings with those of the same population. Dental casts of 46 thalassemic subjects, 25 males and 21 females, aged 7.3 to 23.7 years, were measured for the mesiodistal and buccolingual crown diameters. Crown size variability, correlations, sexual dimorphism, and summary measurements are presented. All means for mesiodistal and buccolingual dimensions in males exceeded those in females, with 18 of the 28 comparisons statistically significant (ranged from P&lt;0.05 to P&lt;0.001, t-test). With the exception of maxillary central and lateral incisors and mandibular first molars, all other teeth exhibited greated buccolingual diameters than mesiodstal. No specific pattern of percentages of sexual dimorphism was noted between the mesiodistal and buccolingual diameters. Comparison of the mesiodistal crown diameters in thalassemic subjects with unaffected control group showed that thalassemic males and females have significantly smaller dimensions than their controls, with 9 of the 24 comparisons being highly significant.

EVALUATION OF HEARING IN PATIENTS WITH BETA THALASSEMIA MAJOR

EVALUATION OF HEARING IN PATIENTS WITH BETA THALASSEMIA MAJOR Ahmed Fadhil Hasan*, Husam Haider Salman# & Jaafar Muhsen Khalaf@ *MB, ChB, FICMS, CABS ENT HNS, ENT Senior Surgeon at Al Shafaa General Hospital. #FRCS, DLO. Retired, Department Of Surgery, College Of Medicine, Basrah University. @MB,ChB, FICMS ENT, ENT Surgeon, Basrah Teaching Hospital, Basrah, IRAQ. Abstract This prospective study was conducted in Basrah Teaching Hospital to evaluate the hearing of β thalassemia major patients, and to find the association between desferoxamine (DFO) therapy and hearing loss (HL). Pure tone eudiometry (PTA) and tympanometry was done to determine hearing thresholds of (23) β thalassemia major patients and (50) non thalassemic subjects. All subjects were more than 4 year old to allow PTA to be done. The criteria of evaluation included a full otological history, drug history, otological examination, audiological evaluation using PTA and tympanometry. This study found that (6) out of (23) thalassemic patients and (6) out of (50) non thalassemic patients have hearing loss. From those thalassemic patients with HL, (5) patients have conductive hearing loss (CHL) and one have sensorineural hearing loss (SNHL), while the non thalassemic patients with HL all have CHL, also we found that (3) out of (5) thalassemic patients not on desferoxamine therapy have HL. In conclusion, β thalassaemia major has no significant statistical association with hearing problems, and there is no significant statistical association between desferoxamine therapy and hearing loss.